# 961: THERMOVIBRIO MEDIUM


## Main sol. 961

0.5	ml	Sodium resazurin (0.1% w/v)	
1	g	KNO3	
1	g	Yeast extract (OXOID)	
0.5	g	Na2S x 9 H2O	
1000	ml	**Synthetic seawater**	

1. Dissolve resazurin in seawater, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into serum vials (e.g., 20 ml medium per 100 ml bottle) and autoclave. Add nitrate, yeast extract and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 6.5, if necessary.
2. After inoculation pressurize bottles to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
3. Note: Alternatively, aged filtered seawater can be used instead of synthetic seawater.


## Synthetic seawater

27.7	g	NaCl	
7	g	MgSO4 x 7 H2O	
5.5	g	MgCl2 x 6 H2O	
0.65	g	KCl	
0.5	g	KH2PO4	
0.25	g	(NH4)2SO4	
0.03	g	H3BO3	
0.1	g	NaBr	
0.75	g	CaCl2 x 2 H2O	
0.05	mg	KI	
15	mg	SrCl2 x 6 H2O	
1000	ml	Distilled water